本试验旨在研究体外培养中硒(Se)对凡纳滨对虾(Litopenaeus vannamei)肝胰腺细胞抗氧化能力的影响以及对黄曲霉毒素B1(AFB1)毒性的缓解作用。在体外培养肝胰腺细胞的基础上,设定含不同浓度(0、0.15、0.30、0.45、0.60、0.75 μg/mL)Se的培养液,研究Se对肝胰腺细胞抗氧化能力的影响;另用浓度为1 600 μg/L的AFB1侵染细胞,探究Se对AFB1毒性的缓解作用。结果表明:当Se浓度处于0~0.45 μg/mL时,随着Se浓度的增加,丙二醛(MDA)含量呈现降低趋势,但各个浓度之间并没有显著差异(P>0.05),过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活力随着Se浓度的增加而增强,Se浓度为0.45 μg/mL时GSH-Px活力显著高于其他浓度(除0.30 μg/mL)时(P<0.05);当Se浓度处于0.45~0.75 μg/mL时,随着MDA含量升高的同时CAT、GSH-Px活力下降。当细胞受到浓度为1 600 μg/L的AFB1侵染后,随着培养时间的增加,MDA含量表现一直升高的趋势,同时CAT、GSH-Px活力都有一定程度的下降;当细胞受到浓度为1 600 μg/L的AFB1和浓度为0.45 μg/mL的Se共同作用后,随着培养时间的增加,MDA含量有一定程度的改善,CAT、GSH-Px活力得到一定程度的恢复。由此可见,在体外培养条件下,浓度为0.45 μg/mL的Se有利于提高凡纳滨对虾肝胰腺细胞的抗氧化能力,且在一定程度上能缓解AFB1对肝胰腺细胞的毒性作用。
The effects of selenium (Se) on antioxidant capacity and relieving toxicity of aflatoxin B1 (AFB1) of Litopenaeus vannamei hepatopancreas cells were evaluated in vitro. A total of 6 concentrations of Se were prepared on the basis of hepatopancreas cells in vitro, which were 0, 0.15, 0.30, 0.45, 0.60 and 0.75 μg/mL, respectively, to evaluate the antioxidant capacity of hepatopancrease cells of shrimp. The same treatment was conducted to the infected cells by 1 600 μg/L AFB1, to observe the relieving toxicity effect of Se. The results showed that malondialdehyde (MDA) content showed a decreased trend with Se concentration increased from 0 to 0.45 μg/mL, but there was no significant difference in all concentrations (P>0.05). The activities of catalase (CAT) and glutathione peroxidase (GSH-Px) were strengthen with the Se element concentration increased from 0 to 0.45 μg/mL, and the activity of GSH-Px in 0.45 μg/mL was significantly higher than in other concentrations except 0.30 μg/mL (P<0.05). As the MDA content increased, the CAT and GSH-Px activities declined, when the concentrations of Se were in 0.45 to 0.75 μg/mL. When cells were infected with 1 600 μg/L AFB1, the content of MDA had a rising trend, and the activities of CAT and GSH-Px had a downward trend with the culture time prolonged. When cells were subjected to the interaction of 1 600 μg/L AFB1 and 0.45 μg/mL Se, the content of MDA had a certain degree of improvement, and the activities of CAT and GSH-Px to restore a certain level with the culture time prolonged. So, 0.45 μg/mL of Se is the optimal concentration, which can improve the antioxidant capacity of Litopenaeus vannamei hepatopancreas cells in vitro. And to a certain extent, 0.45 μg/mL Se can alleviate toxicity effect of AFB1 on hepatopancreas cells in vitro.
[1] 刘永旺,刘海舰,王宝琴,等.富硒麦芽对链尿佐菌素诱发糖尿病大鼠糖脂代谢的影响[J].南京农业大学学报,2004,27(3):81-84.
[2] TAPIERO H,TOWNSEND D M,TEW K D.The antioxidant role of selenium and seleno compound[J].Biomedicine & Pharmacotherapy,2003,57(3/4):134-144.
[3] JULIAN E.Selenium and the prevention of cancer (Part Ⅰ:evidence for the carcinostatie activity of Se compounds)[J].Bulletin of Selenium-Tellurium Development Associatio,2001,5:1-6.
[4] 赵任,郁宝铭,郑民华,等.硒对淋巴细胞杀伤大肠癌细胞过程中Fas/FasL表达的影响[J].中华实验外科杂志,2001,18(1):21-22.
[5] BRIGELIUS-FLOHÉ R,FLOHÉ L.Is there a role of glutathione peroxidases in signaling and differentiation?[J].BioFactors,2003,17(1/2/3/4):93-102.
[6] TAMURA T,STADTMAN T C.A new selenoprotein from human lung adenocarcinoma cells:purification,properties,and thioredoxin reductase activity[J].Proceedings of the National Academy of Sciences of the United States of America,1996,93(3):1006-1011.
[7] SHEN H M,SHI C Y,LEE H P,et al.Aflatoxin B1-induced lipid peroxidation in rat liver[J].Toxicology and Applied Pharmacology,1994,127(1):145-150.
[8] 陈锋,李小霞,潘庆,等.不同硒源对凡纳滨对虾(Litopenaeus vannamei)生长和抗氧化性能的影响[C]//第九届世界华人鱼虾营养学术研讨会论文摘要集.厦门:中国水产学会动物营养与饲料专业委员会,2013:323.
[9] 储霞玲,曹俊明,赵红霞,等.饲料中联合添加硒和谷胱甘肽对凡纳滨对虾生长、饲料系数和体成分的影响[J].饲料工业,2008,29(12):28-31.
[10] 田文静.饲料中添加硒和镁对中华绒螯蟹幼蟹生长、抗氧化性能的影响[D].硕士学位论文.上海:华东师范大学,2014.
[11] 李小霞,陈锋,潘庆,等.酵母硒对凡纳滨对虾生长和抗氧化性能的影响[J].华南农业大学学报,2014,35(6):108-112.
[12] 梁萌青,王家林,常青,等.饲料中硒的添加水平对鲈鱼生长性能及相关酶活性的影响[J].中国水产科学,2006,13(6):1017-1022.
[13] BOONYARATPALIN M,SUPAMATTAYA K,VERAKUNPIRIYA V,et al.Effects of aflatoxin B1 on growth performance,blood components,immune function and histopathological changes in black tiger shrimp (Penaeus monodon Fabricius)[J].Aquaculture Research,2001,32(Suppl.1):388-398.
[14] SMELA M E,CURRIER S S,BAILEY E A,et al.The chemistry and biology of aflatoxin B1:from mutational spectrometry to carcinogenesis[J].Carcinogenesis,2001,22(4):535-545.
[15] MEXíA-SALAZAR A L,HERNÁNDEZ-LíPEZ J,BURGOS-HERNÁNDEZ A,et al.Role of fumonisin B1 on the immune system,histopathology,and muscle proteins of white shrimp (Litopenaeus vannamei)[J].Food Chemistry,2008,110(2):471-479.
[16] 郑丽莉,朱宇旌,邵彩梅,等.动物体内细胞色素酶P3A4与饲料黄曲霉毒素B1毒性的关系[J].动物营养学报,2013,25(1):13-20.
[17] 高玮,黄灿华,兰萍章,等.斑节对虾肝胰腺和血淋巴组织细胞的体外培养[J].中山大学学报:自然科学版,2000,39(S):119-122.
[18] 王荣梅,苏荣胜,潘家强,等.黄曲霉毒素对动物免疫及抗氧化能力的影响[J].饲料研究,2010(2):54-56.
[19] 耿忠诚,王秀娜,王燕,等.不同硒源对仔猪生产性能和抗氧化能力的影响[J].黑龙江八一农垦大学学报,2010,22(6):31-35.
[20] 王宏伟.饲料中的硒和锰对虾体抗氧化酶系统的影响[D].博士学位论文.保定:河北大学,2005.
[21] 韦宇.硒对凡纳滨对虾生长及抗氧化能力的影响[D].硕士学位论文.广州:华南农业大学,2008.
[22] ATROSHI F,BIESE I,SALONIEMI H,et al.Significance of apoptosis and its relationship to antioxidants after ochratoxin a administration in mice[J].Journal of Pharmacy and Pharmaceutical Sciences,2000,3(3):281-291.
[23] CHOI Y K,JUNG K K,CHAE K Y,et al.Effects of vitamin E and selenium supplementation to diets containing aflatoxin B1 on the contents of liver lipids and various blood parameters in rats[J].Asian-Australasian Journal of Animal Science,1995,8(4):379-385.
[24] RASTOGI R,SRICASTABA A K,RASTOGI A K.Long term effect of aflatoxin B1 on lipid peroxidation in rat liver and kidney:effect of picroliv and silymarin[J].Phytotherapy Research,2001,15(4):307-310.
[25] 芮小丽,陈思潭,李春梅.硒对黄曲霉毒素B1暴露肉鸡肝脏氧化损伤的保护作用[J].动物营养学报,2014,26(8):2281-2288.
[26] 王钟翊,李前勇,张德志.纳米硒对肉鸭日粮中黄曲霉毒素B1拮抗作用的研究[J].粮食与饲料工业,2009(9):38-41.
[27] 符晨星,贺建华,侯德兴.硒对动物黄曲霉毒素B1中毒的缓解作用[J].湖南饲料,2011(6):27-32,38.
[28] 郭剑英,郭铭生,潘家强,等.黄曲霉毒素B1、复方中药和硒对肉鸡肝组织匀浆中自由基代谢的影响[J].中国家禽,2012,34(7):19-22.
[29] CHEN K J,FANG J,PENG X.et al.Effect of selenium supplementation on aflatoxin B1-induced histopathological lesions and apoptosis in bursa of Fabricius in broilers[J].Food and Chemical Toxicology,2014,74:91-97.
[30] PARVEEN F,NIZAMANI Z A,GAN F,et al.Protective effect of selenomethionine on aflatoxin B1-induced oxidative stress in MDCK cells[J].Biological Trace Element Research,2014,157(3):266-274.
[31] 冯跃生.硒对黄曲霉毒素B1所致乌骨鸡肝脏损害的保护作用[J].中国兽医科技,1997,27(4):28-29.
京公网安备 11010802041926号